MSc. Project

July 23, 2010

Dissertation Submitted

I’m excited to say I finally finished up writing and have submitted my dissertation on Brettanomyces yeasts titled “Pure Culture Fermentation Characteristics of Brettanomyces Yeast Species and Their Use in the Brewing Industry”. The project took an additional five months of compiling data and writing to get it all finished. In the end, I’m very […]

June 30, 2010

Brettanomyces Yeast Cell Images

While lately I’ve mostly been concentrating on writing and compiling all the information necessary to complete this dissertation, I recently took some time to present a lot of my findings to the Fort Collins Home brew club The Liquid Poets. During this I was able to meet Remi, the yeast wrangler for the home brew […]

December 3, 2009

Data Collection Methods of Analysis

Pure culture fermentations were given 35 days (5 weeks) to attenuate before data and samples were collected and frozen for later analysis. Samples of un-fermented wort had previously been collected and frozen for comparative analysis along with samples from the batch culture propagations. It may be useful to observe the different quantities of compounds produced […]

November 1, 2009

Brettanomyces Research Fermentation Photos

Thought I would include some photos of the pure culture fermentations to give some insight into the research project and show how I set up to observe 105 trial fermentations. CMY001 & BSI-Drie strains along with WhiteLabs strains in a temperature controlled incubator Close up with some of the fermentations initially containing lactic acid CMY001 […]

October 8, 2009

Research Methodology for Pure Culture Fermentation

While performing the propagation trials, I prepared the research methodology for the pure culture fermentations that would make up the bulk of the research. As previously mentioned all fermentations were conducted with wort produced on the 2 hectolitre pilot system. The following details the trials I put together for researching the characteristics of primary fermentation […]

October 1, 2009

Species Identification

Over the past couple of months I’ve been emailing with a graduate student at the University of California, Santa Cruz who was interested in running PCR identification on the two Brettanomyces strains (CMY001 and BSI-Drie) along with a few others. I just received an email detailing that PCR identification sequencing had been carried out on […]

August 8, 2009

MYPG Compared to Wort as a Growth Substrate

While most of the batch culture propagations were conducted using wort as the growth substrate, I also looked at how batch culture growth proceeded using MYPG as the growth substrate. This study was performed with each of the three Brettanomyces strains from Wyeast by inoculating them into 100 ml of MYPG substrate and 100 ml […]

July 27, 2009

Propagation Data and Graphs

I have finished compiling the data on batch culture propagations conducted with five of the Brettanomyces strains. The following two graphs contain data from the standardized propagation method described in the previous post using 12 °Plato (1.048 gravity) wort as the growth medium. Some variation between growth patterns can be seen along with what appears […]

May 15, 2009

Propagations Standardized

During the month of April and beginning of May, I worked to standardize batch culture propagation methods for maximizing cell growth. I had previously finished observing colony morphology and growth characteristics on various mediums and chose MYPG and WLN agar as my two mediums of choice. With eight strains of Brettanomyces to use throughout the […]

May 6, 2009

Wyeast to the Rescue

4/30/09 – Thursday Over the past month or so I have been e-mailing with Greg Doss of Wyeast Labs as previously mentioned and it turns out the gracious people over at Wyeast were able to supply the Brettanomyces Masters Project with three slopes. WY5112 (B. bruxellensis), WY5526 (B. lambicus) and WY5151 (B. claussenii) were shipped […]

April 30, 2009

Media Cultureablity of Brettanomyces Species

Screening of Various Media Agars I have finished collecting and compiling data on the ability of B. bruxellensis (WLP650), B. lambicus (WLP653), B. claussenii (WLP645), B. bruxellensis (BSI-Drie) and B. bruxellensis (CMY001) to culture on popular brewery related media agar. This is the first part of the research and was carried out to find the […]

April 20, 2009

The Many Faces of Brettanomyces

Giant Colony Morphologies When I undertook this project I never imagined the microbiology involved and the hours I would spend in the lab just to get this project off the ground. Three months in the lab culturing organisms so far and I have found that just working with Brettanomyces spp. on various media agars can […]

April 12, 2009

Project Updates

Over the past couple of weeks I have been in talking with Greg Doss the QC manager at Wyeast labs and they will be helping out the Brettanomyces Masters Project by shipping over slopes of B. lambicus (5526), B. bruxellensis (5112), and the VSS for April to June B. claussenii (5151). Thanks to Greg for […]

April 6, 2009

Two for One Brettanomyces Strains

The “Drie” strain(s) As can be seen from reading through the blog, I have been repeatedly culturing what I thought was a single Brettanomyces strain acquired from the dregs of a bottle of Avery’s 15th Anniversary ale. The strain they used for the 100% Brettanomyces fermentation is referred to as “Drie“, as it was cultured […]

March 18, 2009

Day in the Pilot Brewery

Wednesday Pilot Brewing Finally everything has progressed to the point where I can produce a standard 12°Plato (1.048) wort which will be used for each batch propagation and mini-fermentation throughout the Brettanomyces Masters Project. The ICBD pilot brewery is a custom built brewery which produces two hectoliter (200 liter) batch volumes. The grain bill for […]

March 13, 2009

Fun with White Labs

Friday Lab Happenings Finally got a chance to plate out the three White Labs vials which came in last week. As I need to make back-ups of all the Brettanomyces spp. which I collect in order to possibly re-propagate throughout the research, I inoculated two MYPG agar plates for each strain. The plates were inoculated […]

March 6, 2009

WLN Agar Medium

A New Selective Media Agar Wallerstein Laboratory Nutrient (WLN) is a medium which is prepared already and sold by various lab companies. I used WLN from the Oxiod company with an additional 1% (1 g/l) of agar added to help further solidify the media for better streaking ability without gouging into the media. Here is […]

March 5, 2009

White Labs Brettanomyces Strains

White Labs yeasts arrive! Brettanomyces claussenii (WLP645), Brettanomyces bruxellensis (WPL650) and Brettanomyces lambicus (WLP653) have all arrived and will be inoculated onto MYPG agar plates and onto MYPG agar slopes for storage. I will also be propagating up each of the strains to observe growth curves during batch culture propagation. This will be the second […]

February 23, 2009

One Brett So Many Plates

Monday Observations I knew I had made up a lot of plates but when it came to critically observing each plate for growth and deciding how I should follow up with the results, it becomes time consuming and taking good notes is critical. It is counter balanced when observing the aroma of the plates though, […]

February 19, 2009

Inoculation for Pure Culture Confirmation

2/17/09 – Tuesday Inoculation day Using the yeast slurry from each of the four flasks used to propagate up the Avery 15 Brettanomyces sp., I inoculated three different mediums (Copper Sulfate, MYPG+cycloheximide and Lysine agar) prepared the day earlier. First technique was to loop cells from the slurry and streak inoculate each of the different […]

February 16, 2009

Choosing Selective Agar Medias

Monday Lab Prep Made up three different media agars to promote the selective growth of Brettanomyces spp. and use for enumeration. I will test these out with the currently propagating cultures and save them for further use throughout the study to confirm my fermentations are uniquely Brettanomyces spp. The three media agars I chose to […]

February 12, 2009

Propagation Observation

2/10/09 – Tuesday Propagation Observation There was no visible growth in the Brettanomyces propagation flasks yet as each flasks media solution was still unclouded. Further, there was no growth on the MYPG agar plates created from the single colony streaks. I will give it 48 hours and check back again on both. 2/12/09 – Thursday […]

February 9, 2009

Avery 15 Culture Plates

2/6/09 – Friday Observations I observed the plates for the first time after a 48 hour incubation period. There was some barely observable growth, a colony or two on the MYPG plates and one of the MYPG w/CaCO3 plates. No observable smells yet other then the agar media. I left the plates to further incubate […]

February 4, 2009

Inoculating and Culturing the Dregs of Avery 15

Culturing Yeast From Avery’s 15th Anniversary Beer A few days prior to inoculating plates, I removed the special foil (and glue used to hold the foil) from the top and underside of an Avery 15 bottle cap by thoroughly cleaning and scrubbing with warm soapy water. After rinsing and drying the bottle, the neck of […]

February 3, 2009

Lab Preparations

Day 1 in the Lab Made up two additions of MYPG agar in 500 ml volumes. One was with the addition of CaCO3 and the other without CaCO3. MYPG w/CaCO3 is similar to Dekkera media agar recommended for use in culturing Brettanomyces spp. by the European Brewery Convention (EBC). The calcium carbonate is supposed to […]

February 1, 2009

The Beginning of something WILD

The Brettanomyces Masters Project This MSc. dissertation is the result of my enthusiasm and interest in Sour/Wild ales and their unique flavors and aromas. Over the past decade, Brettanomyces spp. have seen an increasing use in the craft-brewing sector of the industry with a handful of breweries having produced beers that were primary fermented with […]